Differential seminal plasma proteome according to semen retrieval in men with spinal cord injury.

OBJECTIVE: To evaluate protein expression profile and to quantify proteins present in seminal plasma from men with spinal cord injury (SCI) and healthy men without SCI. DESIGN: Experimental study. SETTING: University hospital. PATIENT(S): Twelve SCI patients divided into two groups, six who underwent electroejaculation (EEJ) and six who underwent penile vibratory stimulation (PVS); and ten control subjects presenting normal sperm motility and concentration. INTERVENTION(S): EEJ and PVS. MAIN OUTCOME MEASURE(S): The seminal plasma protein profile was analyzed by two proteomic strategies: data-independent label-free quantitative proteomics (MS(E)) and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2D SDS-PAGE). RESULT(S): A total of 638 different proteins were identified by MS(E) and 18 by 2D SDS-PAGE followed by tandem mass spectrometry. Interactome analysis showed key reproductive biologic processes-insemination, sperm and oocyte fusion, and acrosome reaction-related to all groups, as were triglyceride stimuli. Processes related to actin and muscle function and to iron oxidation, transportation, and homeostasis were found only in the EEJ and PVS groups; response to hydrogen peroxide and increased immune response was found only in the PVS group. CONCLUSION(S): This study was able to demonstrate differential protein expression among control, PVS, and EEJ groups; SCI is responsible for alterations in seminal plasma protein profile leading to a deviation from homeostasis; proteins reported in both PVS and EEJ groups correlate with the pathophysiology of SCI-related infertility.

Proteomic analysis of follicular fluid from women with and without endometriosis: new therapeutic targets and biomarkers.

Endometriosis is a gynecological disease that affects women of reproductive age. The protein profiles of women with endometriosis who were able or unable to achieve pregnancy and women without endometriosis who did achieve pregnancy were compared in this study. The follicular fluid was collected from 21 patients undergoing in vitro-fertilization treatment, according to the following groups: nine women in the control group (Group C), four women with endometriosis who achieved pregnancy (Group E.P), and eight women with endometriosis who did not achieve pregnancy (Group E.NP). Follicular fluid proteins were separated using 2D-electrophoresis, and their spots were compared, excised, and submitted to LC-ESI-MS/MS for proteins identification. The analysis showed 29 differentially expressed spots among the groups, and from these, 21 proteins were identified. Analysis showed some functional enrichment in the E.P group, including response to oxidative stress and apoptosis, while the E.NP group showed functions related to response to reactive oxygen species and positive regulation of apoptosis. These data suggest that endometriosis leads to differential protein expression in the follicular fluid, which can influences the outcome of pregnancy. These proteins may be potential targets for better diagnostics and new therapeutic intervention in affected women, as well as assisting in comprehending the physiopathologic mechanisms underlying endometriosis.

Effect of varicocele on sperm function and semen oxidative stress.

OBJECTIVE: To assess the effect of varicocele on sperm DNA integrity, mitochondrial activity, lipid peroxidation and acrosome integrity. PATIENTS AND METHODS: In all, 30 patients with a clinically diagnosed varicocele of grade II or III and 32 men without a varicocele were evaluated for sperm DNA fragmentation (comet assay), mitochondrial activity (3,3'-diaminobenzidine assay), lipid peroxidation (malondialdehyde) and acrosome integrity (fluorescent probe labelled peanut agglutinin). RESULTS: The varicocele group showed fewer spermatozoa with intact DNA (grade II, P= 0.040), more cells with inactive mitochondria (class III, P= 0.001), fewer cells with active mitochondria (class I, P= 0.005) and fewer spermatozoa with intact acrosomes (P < 0.001). Finally, no significant differences were observed in lipid peroxidation levels. CONCLUSION: Men with varicocele showed an increase in sperm DNA fragmentation and a reduction in mitochondrial activity and acrosome integrity. However, lipid peroxidation levels remained unchanged.

Avaliação do espermograma de leões africanos (Panthera leo, Linnaeus, 1758), mantidos na Fundação Parque Zoológico de São Paulo / Spermogram of african lions (Panthera leo, Linnaeus, 1758) housed at Fundação Parque Zoológico de São Paulo

O presente estudo teve por objetivo avaliar o espermograma de um grupo de leões africanos mantidos em cativeiro na Fundação Parque Zoológico de São Paulo, como um aspecto a ser considerado na avaliação do potencial reprodutivo para a aplicação de técnicas dereprodução assistida. Foram submetidos a eletroejaculação 14 exemplares machos, adultos de Leão Africano, utilizando-se o método da eletroejaculação. Logo após a colheita, cada uma das 13 amostras obtidas foi examinada quanto ao volume, pH e aspecto geral, seguida pela avaliação da motilidade, movimento progressivo, índice demotilidade espermática e concentração. Uma alíquota do ejaculado foi fixada em solução de formol-salina a 10% e analisado em câmara úmida ao microscópio de interferência de fase, para o estudo dos aspectos morfológicos, classificando as alterações dos espermatozóides em defeitos maiores e menores. Os resultados foram: volume 5,83 ±3,35 ml; concentração 11,62 ± 14,51 x 106 espermatozóides/ml; motilidade total 73,85 ± 11,02 %; motilidade progressiva 3,35 ±0,63; índice de motilidade espermática 70,42 %, pH 8,1 ± 0,5; defeitos maiores 38,12 ± 19,41 %; defeitos menores 17,43 ± 10,36 %; defeitos totais 55,55 ± 19,30 %. Os defeitos maiores mais freqüentes foram gota citoplasmática proximal, peça intermediária dobrada com gota citoplasmática e peça intermediária dobrada. A cauda dobrada foi o defeito menor mais encontrado.

The aim of this study was to analyse the spermogram of a group of captive African lions (Panthera leo), housed at the Fundação Parque Zoológico de São Paulo, as a subject to avaluate the reproductive potential of the animals, an important aspect to apply artificial reproductive techniques. After chemical restraining, semen from 14 adult male lions was obtained by electroejaculation and the samples were examined right after the collection. The characteristics analyzed were: volume, pH, general aspect, motility, progressive motility, spermatic motility index and concentration. One aliquot of the samples were fixed in saline-formol solution 10% and examined under Phase Contrast Microscopy for morphological evaluation, classifying in minor and major defects. The results were: volume 5,83+/-3,35 ml; concentration11, 62 +/- 14,51 x 106 sperm cells/ml; total motility 73,85 +/- 11,02 % ; progressive motility 3,35 +/- 0,63; spermatic motility index 70,42 %, pH 8,1 +/- 0,5; major defects 38,12+/- 19,41 %; minor defects 17,43 +/- 10,36 %; total defects 55,55 +/- 19,30 %. The more frequent major defects were: proximal droplet, bent midpiece with cytoplasmic droplet and bent midpiece. Bent tailwas the most frequent minor defect observed.